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Table of Content

    28 December 2022, Volume 39 Issue 6
    Analysis of Blood Biochemical Indexes in SPF Beagle Dogs
    HU Jinping, CHEN Jian, ZHANG Jinyu, REN Wenzhi
    2022, 39(6):  1-4.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 001
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    Objective To study and explore the changes of blood biochemical indexes in SPF dogs at different months of age. Method The experiment randomly selected healthy Beagles of different months of age, 12 Beagles of 2, 6 and 9 months of age, and collected 10 mL blood from forelimb vein. The blood biochemical indexes were detected. Result The content of inorganic salts in the blood of SPF Beagles was rich, and the content of Na ions was the most, and the fluctuation was not obvious. Serum alanine aminotransferase (ALT) fluctuated greatly from 2,6, 9 months of age, but there was no significant difference. Total bilirubin ( TBIL) content did not change significantly during the growth of SPF dogs, but remained stable. Although the content of blood glucose and blood lipid fluctuated little during the growth process, it was relatively stable. Conclusion The blood biochemical indices of SPF class Beagles at different months of age fluctuated, and the result of this study will provide basic information for the subsequent breeding and experimental design of SPF class Beagles.
    Metabolomics Study of Joint Synovial Fluid in Beagle Dogs
    LIU Qingping, HUANG Yongping , CHEN Yun , WAN Juan, TIAN Song, HE Zhengming , HU Manli , DING Yi, BAI Lan, ZHANG Xin
    2022, 39(6):  5-14.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 002
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    Objective To analyze and compare metabolomics characteristics of joint synovial fluid between juvenile and adult beagle dogs. Method The joint synovial fluids were collected from 18 juvenile and 16 adult beagle dogs. Samples were subjected to liquid chromatography and mass spectrometry ( LC-MS) -based nontargeted metabolomics analyses. Result Compared with that of adult dogs, male juvenile animals showed higher levels of metabolites related to amino acid biosynthesis, protein digestion and absorption, mineral absorption, and ABC transporter. In female animals, the contents of metabolites related to amino acid, carbohydrate metabolism and ABC transport were higher in juvenile than adult dogs. Conclusion The bone joint development and health are closely related to metabolic events of amino acids, proteins, minerals and carbohydrates.
    Smyd2-knockout Aggravates Pathological Cardiac Hypertrophy Induced by Stress Overload
    LI Wei, SHI Hongjie, ZHOU Yan, HU Yufeng, XIA Hao
    2022, 39(6):  15-19.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 003
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    Objective To investigate the effect of SET and MYND domain-containing protein 2 ( Smyd2 ) gene knockout on the development of pathological cardiac hypertrophy induced by pressure overload. Method Ten wildtype mice and 10 Smyd2 knockout mice were randomly divided into pathological cardiac hypertrophic model group. The pathological cardiac hypertrophy was established by transverse aortic constriction ( TAC) surgery in mice. The structure and function of the heart were detected by echocardiography 4 weeks after surgery, and the degree of cardiac hypertrophy and fibrosis was evaluated by histological pathological staining. Result At 4 weeks after aortic arch ligation, the degree of cardiac hypertrophy and fibrosis in Smyd2 knockout mice was significantly higher than that wild-type mice. Ultrasound result showed that Smyd2 knockout increased the impairment of cardiac function caused by stress load. Conclusion Smyd2 deficiency exacerbates pathological myocardial hypertrophy, fibrosis and heart dysfunction induced by stress load.
    Protective Effects of Deep Electrical Stimulation of the Lateral Cerebellar Nucleus on the Cortical Local Field Potential in a Rat Model of Focal Cerebral Ischemia Induced by Endothelin-1
    SHI Guorong, LIU Tingting, TIAN Xin, ZHU Zixin, ZHENG Wenrong, WANG Yufeng, SUN Fangling, WANG Wen
    2022, 39(6):  20-25.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 004
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    Objective This article aims to investigate the effect of electrical stimulation of the lateral cerebellar nucleus on the LFP in rats with focal cerebral ischemia induced by ET-1. Method Healthy adult male SD rats were injected with ET-1 into the motor cortex to induce a model of focal cerebral ischemia. After 1 week of recovery, rats were installed with self-made stimulation electrode and recording electrode. Then electrical stimulation of 30 HZ were performed for 8 hours per day for 14 consecutive days. Local field potentials evoked during forepaw stimulation were recorded by electrophysiological recorderto analyzed evoked activity of motor cortex. Result After focal ischemia induced by ET-1, the evoked acticivty of the ischemic cortex was significantly reduced, compared with that of the contralateral cortex (P<0. 001) . The induced LFP of the ischemic cortex was significantly increased after electrical stimulation( P < 0. 001) , but still lower than that of the contralateral cortex (P<0. 01) . The result of LFP in all frequency bands showed that the average inter-hemispheric power ratio were increased in Theta band (P<0. 01) , Alpha band (P<0. 05) and beta-1 band (P<0. 01) after DBS for 14 days. Conclusion After ET-1-induced cortical ischemia, average power in LTP was reduced, indicating the decrease of cortical activity, while LCN DBS could improve the excitability of cortical neurons.
    Discussion of Production Technology Produce High-quality Experimental Pigs from Common Sows
    HUA Lizhong, WANG Haiyan, FENG Zhixin, WANG Jia, YUAN Ting, GAN Yuan, ZHANG Lei, SHAO Guoqing
    2022, 39(6):  26-31.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 005
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    Objective To establish the batch production technology of high-quality experimental pigs and realize the batch supply of high-quality experimental animal pigs. Method Two comparative experiments were carried out in this study. Experiment 1: piglets from two erhualian sows and two large white sows were divided into two groups for each litter, and were fed with artificial milk ( experimental group) and commercial infant milk ( control group) respectively; Experiment 2: The piglets from 5 large white sows were selected as the experimental group(5 piglets per litter, fed with artificial milk) and the remaining piglets as the control group fed with breast feeding. Result In experiment 1, (1) the experimental group showed transient diarrhea, and the survival rate at the age of 21 days reached 100%, while the survival rate of the control group was only 10% due to diarrhea. ( 2) All the piglets in this experiment tested antibodies negative for PRRSV, CSFV, PRVgE, PCV2, FMD, and Mhp. The pathogen test also showed negative for PRRSV, PCV2, CSFV, PRV, Mhp, APP, T+Pm, PEDV, and TGEV. In experiment 2, (1) the survival rates of the experimental group and the control group were 100%, and the daily weight gain was 0. 16 and 0. 28 kg / day, respectively, with a significant difference ( p<0. 05) . ( 2) The positive rates of PRRSV, CSFV, PRVgE, PCV2, FMD, and Mhp antibodies were 63. 6%, 84. 8%, 100%, 100%, 100%, and 90. 9% respectively in the control group, and all the other pigs were negative. ( 3 ) Among the above 9 pathogens, PRRSV, PCV2, Mhp and PEDV pathogens in the control group turned positive at weaning, while all indicators in the experimental group were negative. Conclusion Through a barrier system, aseptic delivery, artificial feeding standard, and the optimization of artificial milk formula, a preliminary attempt was developed to artificially breed high-quality experimental piglets with common sows as donors, and high-quality experimental pigs with negative main epidemic pathogens were successfully obtained. This method widens the range of donors and does not require a caesarean section, which is suitable for large-scale promotion and application.
    Genetic Diversity of SD and Wistar Rats was Analyzed by Using 24 Microsatellite Markers
    LI Huan, WEI Jie, ZUO Qin, WANG Hong, ZHOU Jiaqi, GUANG Jiaona, FAN Tao, FU Rui, LIU Zuomin, YUE Bingfei
    2022, 39(6):  32-38.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 006
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    Objective To provide basic data for quality control by analyzing the genetic diversity of SD and Wistar rat populations. Method Using microsatellite DNA labeling technology, 24 microsatellite loci distributed on 21 chromosomes were selected for PCR amplification of DNA samples from 12 pairs of SD rats and Wistar rats, respectively. The amplified products were classified by second-generation sequencing. Then PopGen32 software, Littleprogram0. 6 and GenePop software were used to calculate alleles, average heterozygosity, polymorphism information content and genetic balance test parameters. Result The average number of alleles of 24 microsatellite loci in SD rats and Wistar rats was 3 and 2. 95, respectively. The average heterozygosity was 0. 434 4 and 0. 426 1, and the average polymorphism information content was 0. 38 and 0. 37, respectively. There were 10 and 9 loci deviating from the Hardy-Weinberg equilibrium. Among them, 7 and 4 loci showed heterozygous deletion. Conclusion The genetic diversity of SD and Wistar rat closed populations is slightly lower than that of ideal closed populations. It is necessary to pay attention to scientific management and monitoring in population breeding to avoid inbreeding.
    Effects of Soft Drinks on Weight Gain, Serum Biochemical Indexes and Tissue Cells in Mice
    CAO Yingqi, JIANG Xiaohan, ZENG Huan, LIU Xiaoting, ZHOU Xiangmei, YUAN Jianmin
    2022, 39(6):  39-44.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 007
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    Objective In order to study the effects of drinking soft drinks for a long period of time on the growth, serum biochemical indexes and tissue cells of mice. Method 80 healthy Kunming mice ( half male and half female) were divided into 5 groups, which were free to drink Mirinda, Cola, green tea, coffee and boiled water ( control group) respectively. The other experimental conditions were the same. The body weight, average daily feed intake, serum biochemical indexes, organ coefficient and tissue microscopic changes of mice in different groups were compared. Result The body weights of female mice who drank 4 kinds of soft drinks were higher than the boiled water group in each period. The weights of male mice were significantly higher than the boiled water group (P<0. 05) during 0-50 days, while decreasing during 50-66 days. Serum biochemical indexes indicate that long-term drinking of soft drinks can increase the level of triglyceride in mice (P<0. 05) ; have no significant effect on the organ coefficients of mice (P>0. 05) , but they can cause different levels of degeneration of liver cells and cause protein casts in renal tubules. Conclusion Long-term drinking of soft drinks has a certain effect on the growth and development of mice, and also has certain adverse effects on fat metabolism, liver and kidney function.

    Analysis of the Neural State of Common Raven Calls
    YANG Liqiong, XU Fan, LIU Fangfang
    2022, 39(6):  45-51.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 008
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    Objective The neural or mental states of an organism are distinguished by vocal communication. Method This research used artificial network method ( back propagation neural network ) to classify the neurobiological meaning behind the calls of the common raven ( Corvus corax) . Result There were four types of common raven calls, including alarming, flighting, begging and singing. Artificial intelligence classification can help us distinguish the neural state of animals through the calls of animals. Conclusion Back propagation neural network is a promising candidate method for classification of raven calls.
    Faim2 Gene Knockout Promotes Cardiac Hypertrophy in Mice
    LIU Jiayi , ZHOU Yan, SHI Hongjie, HU Yufeng, LI Hongliang
    2022, 39(6):  52-56.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 009
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    Objective To detect the effect of Faim2 gene knockout on cardiac hypertrophy induced by ransverse aortic constriction ( TAC) in mice. Method Faim2 knockout mice were constructed, and the genotype was identified by PCR and Western blot. Cardiac hypertrophy model was established by TAC. The effects of Faim2 knockout on pathological cardiac hypertrophy were evaluated by hematoxylin-eosin and picrosirius red staining at 4 weeks after TAC surgery. Result Faim2 gene knockout mice were successfully generated. We found that Faim2 knockout could significantly exacerbate cardiac hypertrophy and fibrosis induced by TAC in mice. Conclusion Faim2 gene knockout aggravates cardiac hypertrophy in mice.
    Establishment of Atopic Dermatitis Model Induced by DNCB in BALB / C mice
    ZHENG Xinru, ZHANG Xiaoqing, YANG Yang, WANG Jikui, HAN Xue
    2022, 39(6):  57-62.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 010
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    Objective To establish atopic dermatitis ( AD ) model in BALB / c mice induced by 2, 4- dinitrochlorobenzene (DNCB) , and to explore the optimal conditions, so as to provide theoretical basis for the establishment of animal model in the research of AD. Method BALB / C mice were sensitized and stimulated with different drug concentrations and different modeling time. After the last administration, the severity of dermatitis in the dorsal skin of DNCB-induced AD mice was evaluated based on symptoms, and hematoxylin and eosin ( H&E) and toluidine blue ( TB ) staining were carried out for further analysis. Serum samples were collected for immunoglobulin E ( IgE) detection, and the skin homogenates were prepared to detect the levels of interleukin-4 ( IL-4) , interleukin-5 ( IL-5) and interleukin-13 ( IL-13) . Result Mice in group D were sensitized by 0. 5% DNCB for 3 times and stimulated by 1% DNCB for 6 times. The skin severity score was the highest in group D. The pathological result showed that the skin structure was incomplete, the Spinous layer was obviously thickened, the intracellular edema was obvious, and a large number of inflammatory cells were infiltrated, the levels of serum IgE and cytokines in skin tissue were significantly higher than other groups. Conclusion Low-dose sensitization at early stage and high-dose stimulation at late stage can establish mouse AD model successfully, which lays a foundation for further study on AD.
    Zhuyu Tongmai Capsule alleviates Motor Dysfunction Induced by Middle Cerebral Artery Occlusion (MCAO) and Cerebral Ischemia-reperfusion in Rats
    WANG Jiashuai, LI Xuemei, CAO Chunran, ZHANG Qianqian, ZHU Chunyue, HU Yuchi, GAO Xiaoxin
    2022, 39(6):  63-69.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 011
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    Objective To observe the therapeutic effect of Zhuyu Tongmai Capsule on motor dysfunction induced by middle cerebral artery occlusion ( MCAO) and cerebral ischemia-reperfusion in rats and to study its mechanism. Method The model of middle cerebral artery occlusion ( MCAO) was established by thread occlusion in rats. According to the neural symptom scores of the model animals, the rats were randomly divided into 5 groups after 24 hours: the model group, Zhuyu Tongmai 200, 100, 50 mg / kg group, Ginkgo Leaf Tablets group, and the sham operation group. The rats were administrated by gavage for 14 days. The running test of rats was carried out with gas-2 gait instrument, and the dynamic images of rats’ gait were analyzed. The expression of VEGF, IL-1β, TNF-α in serum was detected by ELISA. Histopathological analysis was carried out in the motor sensory area and hippocampal area of the ischemic side of the brain to observe the neuron damage by HE staining. The expression of nestin and synaptophysin in the ischemic side of the brain was detected by immunohistochemistry. Result Compared with the model group, each dose of Zhuyu Tongmai Capsule can significantly increase the walking speed of the model animals and the average walking cycle; increase the limb stride; reduce the absolute average body angle; reduce four limbs walking cycle,support time and swing time; reduce the braking time and propulsion time of four limbs. On the 14th day after the establishment of the rat model, Zhuyu Tongmai Capsule (200 mg / kg) could enhance the expression of nestin protein in the area around the cortex infarction, but each dose had no significant effect on the content of VEGF, TNF-α and synaptophysin ( SYP ) . Conclusion Zhuyu Tongmai Capsule can significantly promote the recovery of limb motor function in rats with middle cerebral artery occlusion cerebral ischemia, and its mechanism may be related to the enhancement of Nestin protein expression in the area around cerebral infarction, increase of serum VEGF and decrease of serum IL-1β trend,so as to reduce nerve cell injury and promote its neurogenesis of the brain.
    Yinma Jiedu Granule Treats Loperamide-induced Constipation in Mice
    LI Gen, LI Fangyuan, YANG Yang, MA Jingwei, ZANG Caixia, BAO Xiuqi, ZHANG Dan
    2022, 39(6):  70-73.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 012
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    Objective The present study aims to study the therapeutic effect of Yinma Jiedu Granule on loperamideinduced constipation in mice. Method Male ICR mice were randomly divided into control group, model group and Yinma Jiedu Granule 500, 1 000 and 2 000 mg / kg groups. Mice were administrated with 500, 1 000 and 2 000 mg / kg of Yinma Jiedu Granule by gavage each day for continuous 8 days. Thirty minutes after the last administration, loperamide was orally given to induce constipation. Then the ink was intragastric administrated 30 minutes later. The large intestinal propulsion rate, total intestinal propulsion rate, number of black stools in the large intestine, the time of first black stool defecation and the number of defecation in an hour were recorded in each group of mice. Result Yinma Jiedu Granule 2 000 mg / kg could significantly increase the large intestinal propulsion rate and total intestinal propulsion rate, decrease the time of first black stool defecation, and increase the number of defecation within 1 hour compared with constipation of mice induced by loperamide. Conclusion Yinma Jiedu Granule could alleviate loperamide-induced constipation though improving the function of large intestine propulsion, shortening defecation time and increasing defecation volume in mice, suggesting that Yinma Jiedu Granule might be an effective drug for constipation treatment.

    Prokaryotic Expression and Immune Efficacy of OmpA of Rodentibacter heylii
    XING Jin, FENG Yufang, GAO Qiang, ZHANG Xueqing, ZHAO Deming, YUE Bingfei
    2022, 39(6):  74-80.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 013
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    Objective To explore the immune effect of the outer membrane protein A ( OmpA ) prokaryotic expression protein of Rodentibacter heylii ( Rh ) on mice. Method Prokaryotic expression and purification by nickel ion affinity chromatography were used to prepare Rh OmpA recombinant protein, and the target protein was identified by SDS-PAGE and Western blot. The obtained OmpA target protein was immunized to ICR mice by intraperitoneal and intramuscular injection and nasal drip. Subsequently, the antibody titer was determined by ELISA method . Then Rh ATCC12555, Rodentibacter pneumotropicus (Rp) ATCC35149, and Rh isolate PP320 were challenged to the immunized mice abdominal cavity to evaluate the protective effect of recombinant protein OmpA on mice. Result The OmpA specific antibody level in serum of immunized mice was determined by ELISA with the expression protein OmpA as the coated antigen. The average antibody titer was 19. 84log2. The protective rates of immunized mice against two standard strains and PP320 were 60%, 60% and 80%, respectively. The mortality rate was significantly reduced(P<0. 001) . The ELISA method based on OmpA expressed protein detects the sera of Pasteurella pneumotropica( Pp) positive mice, and the antibody positive rate is 60%. Conclusion The prepared Rh OmpA expressed protein had good immunogenicity and had a certain protective effect on Rh and Rp, which can provide a reference for the prevention and control of respiratory pathogens in laboratory animals.
    Precise Nutrition Application in the Feed Formula of Laboratory Animals
    LI Huixian
    2022, 39(6):  81-84.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 014
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    With the rapid development of laboratory animals science, there are more and more species and strains of laboratory animals, which are used in different life research topics, and their demand for nutrition is gradually diversified. Therefore, the precise breeding of laboratory animals is becoming more and more important. It can provide accurate nutritional requirements for laboratory animals, maximize the transfer out of experimental result, and promote the development of life science. This paper mainly describes the application of precision nutrition in the feed formula of laboratory animals.
    Focus of Qualified Supplier Evaluation in Animal Laboratory
    SUN Chenyang, WANG Lei, XU Hufeng
    2022, 39(6):  85-90.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 015
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    With the continuous improvement of modern science and technology, experimental animals have gradually become an indispensable “ living reagent” in the fields of medicine, life science, medicine and other disciplines. The quality of laboratory animals, the supplies required by laboratory animals and the supplies required by the daily work of the laboratory animal center directly affect the scientificity, accuracy and reliability of scientific experiment result . Therefore, it is very important to establish qualified suppliers for laboratories in animal laboratories to ensure the quality of experimental animals, laboratory supplies and scientific experiment result .
    Research Progress on Mice Model of Cholesterol Gallstone and the Design and Application Status of Stone Feed
    SU Yongteng, GE Quanjin, WU Xiaoma
    2022, 39(6):  91-95.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 016
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    Cholelithiasis is a common disease of biliary system which seriously threatens human health. The establishment of animal model of cholelithiasis can help researchers to further study the pathophysiology of the disease and develop innovative and effective therapeutic method . Mice is one of the good animal models for gallstone because of its simple modeling method, fast gallstone formation, high incidence of gallstone in treatment group, and similar composition of gallstone to human. This paper briefly describes the definition, classification and causes of gallstones, summarizes the research progress of animal and mice gallstone models, and summarizes the design and application status of stone feed, so as to provide certain method and data reference for the early research of gallstones.

    Cold Atmospheric Plasma and its Research Progress in Promoting Skin Wound Healing
    LIU Yang, GUO Peng, ZHANG Nan, WANG Fujin, WANG Jingyu, WANG Aiguo
    2022, 39(6):  96-103.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 06. 017
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    Skin wound healing has always been the focus of the medical community. Since the 21st century, the rapid development of cold atmospheric plasma ( CAP ) medicine has provided an excellent treatment method for promoting the healing of skin wound. In this review, the CAP related concept and classification, generation and use method, and its application in medicine is briefly summarized. Further, the relevant molecular mechanisms of CAP in promoting wound healing phases including coagulation, inflammation, migration / proliferation, and remodeling are emphatically reviewed.